National Health and Nutrition Examination Survey

2011-2012 Data Documentation, Codebook, and Frequencies

Human Papillomavirus (HPV) - Oral Rinse (ORHPV_G)

Data File: ORHPV_G.xpt

First Published:
February 2014

Last Revised:
NA

Component Description

The human papillomavirus (HPV) in oral rinse estimates the prevalence and determinants of oral HPV infection in the United States population. Oral HPV infection is newly appreciated as a strong risk factor for a distinct type of oropharyngeal squamous cell carcinoma that is rising in incidence in the United States. It has been estimated that oral HPV16 infection confers an approximate 15-fold increase in risk for oropharyngeal cancer. Despite these strong risks, little is known about the epidemiology of oral HPV infection. No population-based surveys of oral HPV infection, type distribution and determinants have been performed in any population worldwide.

Eligible Sample

Examined participants, aged 14-69 years, were tested. The public data file includes data for persons 18-69 years of age. Please see Analytic Notes about the release of data for adolescents 14-17 years of age.

Description of Laboratory Methodology

Oral rinse specimens were processed, stored and shipped to the Ohio State University Gillison Laboratory, Columbus, Ohio. Detailed specimen collection and processing instructions are discussed in the NHANES Laboratory/Medical Technologists Procedures Manual (LPM).

Samples will be centrifuged at 4,000 x g for 10 minutes at 4°C, the pellet will be resuspended in 1 ml of Puregene cell lysis solution and incubated at 37°C for 15 min followed by DNase-free RNase A (5µg/ml) digestion for 30 min at 37°C. Proteinase K (20 mg/ml) digestion will be performed overnight at 55°C followed by heat inactivation for 10 min at 95°C. Protein precipitation solution (340 µL) will be added and sample will be further purified according to the Puregene DNA purification kit protocol.

Purified DNA will be analyzed for 37 types of HPV by means of a multiplex polymerase-chain-reaction (PCR) assay targeted to the conserved L1 region of the viral genome, using PGYM09/11 primer pools and primers for ß-globin (33). PCR products will be denatured in 0.13 N NaOH and hybridized to an immobilized HPV probe array (Roche Diagnostics) using an extended line-blot assay for genotyping of 37 HPV types, including HPV-6, -11, -16, -18, -26, -31, -33, -35, -39, -40, -42, -45, -51, -52, -53, -54, -55, -56, -57, -58, -59, -61, -62, -64, -66, -67, -68, -69, -70, -71, -72, -73, -81, -82 (MM4 and IS39 subtypes), -83, -84 and -89, and ß-globin (Roche Molecular Systems, Inc., Alameda, CA). Positive controls, consisting of 10 and 100 HPV-16 (SiHa) or HPV-18 (C4–2)-positive cells diluted in a background of HPV-negative cells (K562), and a negative control (K562 cells), in addition to the manufacturer’s controls, are included in each experiment. Samples positive for ß-globin will be considered of sufficient quality for analysis. The HPV type will be reported for positive samples.

Data Processing and Editing

The data were reviewed. Incomplete data or improbable values were sent to the performing laboratory for confirmation.

Analytic Notes

The analysis of NHANES 2011-2012 laboratory data must be conducted using the appropriate survey design and demographic variables. The NHANES 2011-2012 Demographics File contains demographic data, health indicators, and other related information collected during household interviews as well as the sample weight variables. The Fasting Questionnaire File includes auxiliary information such as fasting status, the time of venipuncture, and the conditions precluding venipuncture. The demographics and fasting questionnaire files may be linked to the laboratory data file using the unique survey participant identifier (i.e., SEQN).

Exam sample weights should be used for analyses. Please refer to the NHANES Analytic Guidelines and the on-line NHANES Tutorial for further details on the use of sample weights and other analytic issues.

The public release data file includes Oral HPV data for participants aged 18–69. Data for youth aged 14–17 years are available through the NCHS Research Data Center (RDC).